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Literature summary for 3.4.22.B62 extracted from

  • Corvo, I.; Cancela, M.; Cappetta, M.; Pi-Denis, N.; Tort, J.F.; Roche, L.
    The major cathepsin L secreted by the invasive juvenile Fasciola hepatica prefers proline in the S2 subsite and can cleave collagen (2009), Mol. Biochem. Parasitol., 167, 41-47.
    View publication on PubMed
Search for more literature for 3.4.22.B62

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Hansenula polymorpha strain NCYC 495 Fasciola hepatica
the proenzyme is expressed in the methylotrophic yeast Hansenula polymorpha and the mature enzyme is obtained from the culture medium Fasciola hepatica

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0041
-
 tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin
-
 Fasciola hepatica
0.0041
-
 tosyl-Gly-L-Pro-L-Arg-7-amido-4-methylcoumarin in 50 mM sodium phosphate, pH 6.5, 1 mM dithiothreitol, 1 mM EDTA, at 24°C Fasciola hepatica
0.0157
-
 benzyloxycarbonyl-Val-Leu-Lys-7-amido-4-methylcoumarin
-
 Fasciola hepatica
0.0157
-
 benzyloxycarbonyl-L-Val-L-Leu-L-Lys-7-amido-4-methylcoumarin in 50 mM sodium phosphate, pH 6.5, 1 mM dithiothreitol, 1 mM EDTA, at 24°C Fasciola hepatica
0.0235
-
 benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin
-
 Fasciola hepatica
0.02353
-
 benzyloxycarbonyl-L-Phe-L-Arg-7-amido-4-methylcoumarin in 50 mM sodium phosphate, pH 6.5, 1 mM dithiothreitol, 1 mM EDTA, at 24°C Fasciola hepatica

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
additional information Fasciola hepatica cathepsin L3 is a main component of the juvenile secretory products and may participate in the tissue invasion process ?
-
 ?
additional information Fasciola hepatica does not cleave immunoglobulin ?
-
 ?
Type I collagen + H2O Fasciola hepatica CL3 exhibits the ability to degrade type I collagen efficiently at pH 5.5, 6.3 and 7.0 ?
-
 ?

Organism

Organism UniProt Comment Textmining
Fasciola hepatica
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Fasciola hepatica
HiTrap QXL column chromatography, gel filtration Fasciola hepatica

Source Tissue

Source Tissue Comment Organism Textmining
juvenile cathepsin L3 is a main component of the juvenile secretory products and may participate in invasion Fasciola hepatica
-
larva CL3 is a main component of the juvenile secretory products Fasciola hepatica
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
benzyloxycarbonyl-L-Phe-L-Arg-7-amido-4-methylcoumarin + H2O
-
 Fasciola hepatica benzyloxycarbonyl-L-Phe-L-Arg + 7-amino-4-methylcoumarin
-
 ?
benzyloxycarbonyl-L-Val-L-Leu-L-Lys-7-amido-4-methylcoumarin + H2O
-
 Fasciola hepatica benzyloxycarbonyl-L-Val-L-Leu-L-Lys + 7-amino-4-methylcoumarin
-
 ?
benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin + H2O
-
 Fasciola hepatica benzyloxycarbonyl-Phe-Arg + 7-amino-4-methylcoumarin
-
 ?
benzyloxycarbonyl-Val-Leu-Lys-7-amido-4-methylcoumarin + H2O
-
 Fasciola hepatica benzyloxycarbonyl-Val-Leu-Lys + 7-amino-4-methylcoumarin
-
 ?
Collagen + H2O FhCL3 efficiently cleaves type I collagen over different pH and temperature conditions Fasciola hepatica ?
-
 ?
additional information cathepsin L3 is a main component of the juvenile secretory products and may participate in the tissue invasion process Fasciola hepatica ?
-
 ?
additional information no cleavage of immunoglobulin Fasciola hepatica ?
-
 ?
additional information does not cleave immunoglobulin Fasciola hepatica ?
-
 ?
additional information cathepsin L3 has preference for proline in the selective P2 position Fasciola hepatica ?
-
 ?
tosyl-Gly-L-Pro-L-Arg-7-amido-4-methylcoumarin + H2O restricted substrate specificity with 70fold preference for tosyl-Gly-L-Pro-L-Arg-7-amido-4-methylcoumarin over benzyloxycarbonyl-L-Phe-L-Arg-7-amido-4-methylcoumarin and benzyloxycarbonyl-L-Val-L-Leu-L-Lys-7-amido-4-methylcoumarin Fasciola hepatica ?
-
 ?
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin + H2O 70fold preference for tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin over typical cathepsin substrates with hydrophobic or aliphatic residues in the S2 position Fasciola hepatica tosyl-Gly-Pro-Arg + 7-amino-4-methylcoumarin
-
 ?
Type I collagen + H2O CL3 exhibits the ability to degrade type I collagen efficiently at pH 5.5, 6.3 and 7.0 Fasciola hepatica ?
-
 ?

Synonyms

Synonyms Comment Organism
cathepsin L3
-
 Fasciola hepatica
CL3
-
 Fasciola hepatica
FhCL3
-
 Fasciola hepatica

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.33
-
 benzyloxycarbonyl-Val-Leu-Lys-7-amido-4-methylcoumarin
-
 Fasciola hepatica
0.33
-
 benzyloxycarbonyl-L-Val-L-Leu-L-Lys-7-amido-4-methylcoumarin in 50 mM sodium phosphate, pH 6.5, 1 mM dithiothreitol, 1 mM EDTA, at 24°C Fasciola hepatica
0.39
-
 benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin
-
 Fasciola hepatica
0.39
-
 benzyloxycarbonyl-L-Phe-L-Arg-7-amido-4-methylcoumarin in 50 mM sodium phosphate, pH 6.5, 1 mM dithiothreitol, 1 mM EDTA, at 24°C Fasciola hepatica
6
-
 tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin
-
 Fasciola hepatica
6
-
 tosyl-Gly-L-Pro-L-Arg-7-amido-4-methylcoumarin in 50 mM sodium phosphate, pH 6.5, 1 mM dithiothreitol, 1 mM EDTA, at 24°C Fasciola hepatica

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6
-
 using benzyloxycarbonyl-L-Val-L-Leu-L-Lys-7-amido-4-methylcoumarin as substrate Fasciola hepatica
7
-
-
 Fasciola hepatica
7
-
 using tosyl-Gly-L-Pro-L-Arg-7-amido-4-methylcoumarin as substrate Fasciola hepatica

pH Range

pH Minimum pH Maximum Comment Organism
5 9 CL3 has about 70% of the maximum activity at pH 5.0 and about 40% of the maximum activity at pH 9.0 using benzyloxycarbonyl-L-Val-L-Leu-L-Lys-7-amido-4-methylcoumarin as substrate Fasciola hepatica
5.5 8 CL3 has 35% of the maximum activity at pH 5.5 and about 70% of the maximum activity at pH 8.0 using tosyl-Gly-L-Pro-L-Arg-7-amido-4-methylcoumarin as substrate Fasciola hepatica
6 8 pH 6: about 70% of maximal activity, pH 8.0: abput 70% of maximal activity Fasciola hepatica

pH Stability

pH Stability pH Stability Maximum Comment Organism
7
-
 37°C, 50% loss of activity after 8 h, 64% loss of activity after 24 h Fasciola hepatica
7
-
 50% of the activity is conserved after 8 h incubation and 36% of it is retained after 24 h incubation at pH 7.0 and 37°C Fasciola hepatica

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
16.78
-
 benzyloxycarbonyl-L-Phe-L-Arg-7-amido-4-methylcoumarin in 50 mM sodium phosphate, pH 6.5, 1 mM dithiothreitol, 1 mM EDTA, at 24°C Fasciola hepatica
20.76
-
 benzyloxycarbonyl-L-Val-L-Leu-L-Lys-7-amido-4-methylcoumarin in 50 mM sodium phosphate, pH 6.5, 1 mM dithiothreitol, 1 mM EDTA, at 24°C Fasciola hepatica
1464
-
 tosyl-Gly-L-Pro-L-Arg-7-amido-4-methylcoumarin in 50 mM sodium phosphate, pH 6.5, 1 mM dithiothreitol, 1 mM EDTA, at 24°C Fasciola hepatica